KR20220024325A - Predicting or Diagnosing Composition for Risk of Renal Diseases Using Human Intestinal Microbiome, Diagnosing Kit, Method For Providing Information, And Screening Method For Drugs For Preventing Or Treating Renal Diseases Using The Same - Google Patents

Abstract

The present invention relates to: a composition for predicting or diagnosing the risk of renal diseases using intestinal microbes; a kit for predicting or diagnosing the risk of renal diseases including the same; an information providing method for predicting or diagnosing the risk of renal diseases; and a method for screening an agent for preventing or treating renal diseases. The present invention is intended to predict or diagnose the risk of renal diseases and screen an agent for preventing or treating renal diseases through a formulation capable of detecting one or more strains selected from the group consisting of a Lachnospira sp. strain and Dorea formicigenerans sp. strain, a formulation capable of detecting one or more strains selected from the group consisting of a Bacteroides sp. strain, a Bifidobacterium adolescentis sp. strain, a Bacteroides uniformis sp. strain, or a formulation capable of detecting one or more strains selected from the group consisting of a Roseburia sp. strain, an Oscillospira sp. strain, and a Lachnospiraceae sp. strain.

Description

Predicting or Diagnosing Composition for Risk of Renal Diseases Using Human Intestinal Microbiome, Diagnosing Kit, Method For Providing Information, And Screening Method For Drugs For Preventing Or Treating Renal Diseases Using The Same}

The present invention relates to a composition for predicting or diagnosing renal disease risk using intestinal microorganisms, a diagnostic kit for predicting or diagnosing renal disease risk using the same, a method for predicting or diagnosing renal disease risk using the same, and a method for preventing or screening a renal disease using the same.

With the development of next-generation sequencing techniques, it has become possible to quickly and accurately measure the types and relative amounts of microorganisms living in an individual's gut. In particular, this technology makes it possible to find out the type and relative amount of microorganisms living in an individual's intestines using the 16S ribosomal RNA gene variable region sequence, which is a conserved region common to intestinal microbes, and the information can be used as personal disease information or information. Research is being carried out to determine the association with biomarkers related to disease information.

Korea Patent No. 1940424 describes a method for diagnosing renal disease through metagenome analysis using extracellular vesicles secreted from bacteria from blood or urine samples as a sample, but in that it is derived from blood or urine, not intestinal microorganisms. If there is a difference and the content of vesicles derived from Anaerococcus and Burkholderia ) bacteria is increased, and when the content of vesicles derived from bacteria of the genus Acinetobacter and Bacteroides is increased If it is decreased, it is stated that renal failure is diagnosed.

Chinese Patent No. 110878349 describes that Bacteroides cellulolyticus, Bacteroides fragilis, and Roseburia intestinalis can be used in the diagnosis of kidney disease-related diseases.

Korea Patent No. 1940445 describes that Bacteroides, Pribotella and Luminococcus are highly distributed in the case of kidney disease.

However, the results presented in the above patents are biomarker microorganisms that allow to distinguish between the high-risk group for renal disease and the normal group, the combination of the microorganisms, the characteristic nucleotide sequence for identifying the microorganisms, and the increase/decrease pattern of microorganisms in the high-risk group In the present invention, there was a difference from the research results conducted by the present inventors who identified the association with renal disease based on the microbiome obtained from 890 Koreans, and therefore the present inventors completed the present invention based on the results of this study.

Korea Registered Patent No. 1940424 Chinese Patent No. 110878349 Korean Patent No. 1940445

The present invention includes an agent capable of detecting any one or more strains selected from the group consisting of Lachnospira sp. strain and Dorea formicigenerans sp. strain, renal disease risk using intestinal microorganisms To provide a predictive or diagnostic composition.

Another object of the present invention is to provide a kit for predicting or diagnosing renal disease risk, including a composition for predicting or diagnosing renal disease risk using the intestinal microorganism.

In addition, the present invention detects any one or more strains selected from the group consisting of Lachnospira sp . strains and Dorea formicigenerans sp. strains from genomic DNA of microorganisms obtained from a sample derived from the intestinal tract of a test subject. to do; And in the genomic DNA of the microorganism obtained from the sample derived from the intestinal tract of the testee, Lachnospira spp. strain and Dorea formicigenerans ) The increase or decrease of any one or more strains selected from the group consisting of strains normal group Compared with, in the genomic DNA of the microorganism or Lachnospira , when the spp. strain is reduced, or Dorea formicigenerans ) When the strain is increased, the test subject is tested for renal disease or Diagnosing the renal disease risk group; to provide an information providing method for predicting or diagnosing renal disease risk using intestinal microorganisms, including.

In addition, the present invention Lachnospira ( Lachnospira ) sp. strain, Dorea formicigenerans ) sp. strain, Bacteroides ( Bacteroides ) sp. strain, Bifidobac from genomic DNA of microorganisms obtained from a sample derived from the intestinal tract of a test subject Therium adoles centis ( Bifidobacterium adolescentis ) spp. strain and Bacteroides uniformis spp. strain, Roseburia spp. strain, Oscillospira spp. strain and Lachnospiraceae detecting the relative amount of two or more strains selected from the family and strains; And the result obtained in the detecting step, calculating the obesity risk of the test subject by a multivariate linear model predicting a renal disease group using the relative amount of the two or more strains as a variable; It is to provide an information providing method for predicting or diagnosing the risk of renal disease.

In addition, the present invention Lachnospira ( Lachnospira ) sp. strain, Dorea formicigenerans ) sp. strain, Bacteroides ( Bacteroides ) sp. strain, Bifidobac from genomic DNA of microorganisms obtained from a sample derived from the intestinal tract of a test subject Therium adoles centis ( Bifidobacterium adolescentis ) spp. strain and Bacteroides uniformis spp. strain, Roseburia spp. strain, Oscillospira spp. strain and Lachnospiraceae and detecting the strain; And in the genomic DNA of the microorganism obtained from the sample derived from the intestinal tract of the test subject, Dorea formicigenerans spp. strain, Roseburia spp . strain and Oscillospira sp. strain is increased, or Lachnospira sp. strain, Bacteroides sp. strain, Bifidobacterium adolescentis sp. strain, Bacteroides uniformis sp. strain, and Lachnospira bird ( Lachnospiraceae ) and diagnosing the test subject as a renal disease or renal disease risk group when the strain is reduced; to provide an information providing method for predicting or diagnosing renal disease risk using intestinal microorganisms, including.

In addition, the present invention comprises the steps of administering a candidate substance for the prevention or treatment of renal disease to a non-human animal; Any one or more strains selected from the group consisting of Lachnospira sp . strains and Dorea formicigenerans sp. strains from the genomic DNA of microorganisms obtained from the intestinal-derived sample before and after the candidate material treatment detecting; And by comparing the increase or decrease of any one or more strains selected from the group consisting of Lachnospira sp . strains and Dorea formicigenerans sp. strains in the genomic DNA of microorganisms before and after treatment with the candidate material , Veillonella dispar ( Veillonella dispar ) When the strain is reduced, or Lachnospira ( Lachnospira ) When the genus strain is increased, or Dorea formicigenerans ( Dorea formicigenerans ) The strain is reduced In this case, to provide a screening method for preventing or treating a renal disease using an intestinal microorganism; determining the candidate substance as a preventive or therapeutic agent for renal disease.

The present invention includes an agent capable of detecting any one or more strains selected from the group consisting of Lachnospira sp. strain and Dorea formicigenerans sp. strain, renal disease risk using intestinal microorganisms It relates to a composition for prediction or diagnosis.

The present invention also relates to a kit for predicting or diagnosing renal disease risk, comprising the composition for predicting or diagnosing renal disease risk using the intestinal microbe.

In addition, the present invention detects any one or more strains selected from the group consisting of Lachnospira sp . strains and Dorea formicigenerans sp. strains from genomic DNA of microorganisms obtained from a sample derived from the intestinal tract of a test subject. to do; And in the genomic DNA of the microorganism obtained from the sample derived from the intestinal tract of the testee, Lachnospira spp. strain and Dorea formicigenerans ) The increase or decrease of any one or more strains selected from the group consisting of strains normal group In comparison with, Lachnospira in the genomic DNA of the microorganism is reduced, or when the Dorea formicigenerans species strain is increased, the test subject is renal disease or renal disease It relates to a method of providing information for predicting or diagnosing the risk of renal disease using intestinal microbes, including the step of diagnosing the risk group.

In addition, the present invention Lachnospira ( Lachnospira ) sp. strain, Dorea formicigenerans ) sp. strain, Bacteroides ( Bacteroides ) sp. strain, Bifidobac from genomic DNA of microorganisms obtained from a sample derived from the intestinal tract of a test subject Therium adoles centis ( Bifidobacterium adolescentis ) spp. strain and Bacteroides uniformis spp. strain, Roseburia spp. strain, Oscillospira spp. strain and Lachnospiraceae detecting the relative amount of two or more strains selected from the family and strains; And the result obtained in the detecting step, calculating the obesity risk of the test subject by a multivariate linear model predicting a renal disease group using the relative amount of the two or more strains as a variable; It relates to a method of providing information for predicting or diagnosing renal disease risk.

In addition, the present invention Lachnospira ( Lachnospira ) sp. strain, Dorea formicigenerans ) sp. strain, Bacteroides ( Bacteroides ) sp. strain, Bifidobac from genomic DNA of microorganisms obtained from a sample derived from the intestinal tract of a test subject Therium adoles centis ( Bifidobacterium adolescentis ) spp. strain and Bacteroides uniformis spp. strain, Roseburia spp. strain, Oscillospira spp. strain and Lachnospiraceae and detecting the strain; And in the genomic DNA of the microorganism obtained from the sample derived from the intestinal tract of the test subject, Dorea formicigenerans spp. strain, Roseburia spp . strain and Oscillospira sp. strain is increased, or Lachnospira sp. strain, Bacteroides sp. strain, Bifidobacterium adolescentis sp. strain, Bacteroides uniformis sp. strain, and Lachnospira bird ( Lachnospiraceae ) and the step of diagnosing the test subject as a renal disease or renal disease risk group when the strain is reduced; to a method for providing information for predicting or diagnosing renal disease risk using intestinal microorganisms, including.

In addition, the present invention comprises the steps of administering a candidate substance for the prevention or treatment of renal disease to a non-human animal; Any one or more strains selected from the group consisting of Lachnospira sp . strains and Dorea formicigenerans sp. strains from the genomic DNA of microorganisms obtained from intestinal-derived samples before and after treatment with the candidate material detecting; And by comparing the increase or decrease of any one or more strains selected from the group consisting of Lachnospira sp . strains and Dorea formicigenerans sp. strains in the genomic DNA of microorganisms before and after treatment with the candidate material , Lachnospira ( Lachnospira ) When the sp. strain is increased, or when the Dorea formicigenerans strain is reduced, determining the candidate material as a renal disease prevention or treatment agent; including; It relates to a screening method for preventing or treating renal disease using intestinal microorganisms.

In the present invention, based on the body mass index (BMI) of 890 Korean subjects, those of less than 18.5 were classified into the underweight group, those with more than 18.5 and less than 25 were classified into the normal group, and those with more than 25 were classified into the renal disease group, and the Microorganisms whose average ratio decreases or increases in the order of renal disease group, normal group, and low weight group are searched for, and among the discovered microorganisms, microorganisms showing a statistically significant difference in ratio between the renal disease group and the normal group are used for predicting or diagnosing the risk of renal disease. It was identified as a biomarker strain for

As a biomarker strain for predicting or diagnosing the risk of renal disease showing a statistically significant difference between the renal disease group and the normal group, Veillonella dispar species strain, Lachnospira sp. strain, Dorea Formicigenerans ( Dorea formicigenerans ) species strain, Bacteroides genus strain, Bifidobacterium adolescentis ( Bifidobacterium adolescentis ) species strain, Bacteroides uniformis spp. strain, Roseburia ( Roseburia ) sp. strain, Oscillospira sp. strain and Lachnospiraceae and any one selected from the group consisting of strains or a combination of two or more strains.

By comparing the increase or decrease of the strain, the increase or decrease of the strain means an increase or decrease in the relative ratio of the biomarker strains in the genomic DNA of the microorganism obtained from the sample derived from the test subject's intestinal tract, or the renal disease group and the normal group may mean an increase or decrease in the number of bacteria of the biomarker strains or an absolute value indicating it in can hold it.

Among the biomarker strains, Veillonella dispar ( Veillonella dispar ) strain, preferably Veillonella dispar identified by the 16S rRNA nucleotide sequence of SEQ ID NO: 1, is a new disease group compared to the normal group is significantly higher in Therefore, when the Veillonella dispar strain is higher than that of the normal group in the genomic DNA of the microorganism obtained from the sample derived from the test subject's intestinal tract, it can be predicted or diagnosed as having a high risk of renal disease. In addition, Veillonella dispar ( Veillonella dispar ) When the strain is low compared to the renal disease group, it is predicted or diagnosed as a normal group, so that the renal disease risk can be predicted or diagnosed as low.

Among the biomarker strains, Lachnospira sp. strain, preferably Lachnospira identified by the 16S rRNA nucleotide sequence of SEQ ID NO: 2, is significantly lower in the renal disease group than in the normal group. . Therefore, if the genomic DNA of the microorganism obtained from the sample derived from the test subject's intestinal tract is Lachnospira , when the spp. strain is lower than that of the normal group, it can be predicted or diagnosed as having a high risk of renal disease. In addition, if the Lachnospira spp . strain is higher than the renal disease group, it can be predicted or diagnosed as a normal group and thus predicted or diagnosed as having a low renal disease risk.

Among the biomarker strains, Rhea formicigenerans spp . strain, preferably Dorea formicigenerans , identified by the 16S rRNA nucleotide sequence of SEQ ID NO: 3, the spp. strain is renal disease compared to the normal group was significantly higher in the group. Therefore, in the genomic DNA of the microorganism obtained from the sample derived from the test subject's intestinal tract, if the Dorea formicigenerans species strain is higher than that of the normal group, the risk of renal disease can be predicted or diagnosed. In addition, when the Dorea formicigenerans species strain is lower than the renal disease group, the risk of renal disease can be predicted or diagnosed as low.

Among the biomarker strains, Bacteroides sp. strain, preferably Bacteroides sp. strain identified by the 16S rRNA sequence of SEQ ID NO: 4, is significantly lower in the renal disease group than in the normal group. . Therefore, when the genomic DNA of the microorganism obtained from the sample derived from the test subject's intestinal tract is lower than the normal group, the Bacteroides genus strain can be predicted or diagnosed as having a high risk of renal disease. In addition, if the Bacteroides genus strain is higher than the renal disease group, it can be predicted or diagnosed as a normal group and thus predicted or diagnosed as having a low renal disease risk.

Among the biomarker strains, Bifidobacterium adolescentis species strain, preferably SEQ ID NO: 5 and SEQ ID NO: 6 Bifidobacterium adol identified by any one sequence selected from the rRNA sequence of SEQ ID NO: Rescentis ( Bifidobacterium adolescentis ) strains appear significantly lower in the renal disease group than in the normal group. Therefore, in the genomic DNA of microorganisms obtained from the sample derived from the intestinal tract of the subject, when the Bifidobacterium adolescentis species strain is lower than that of the normal group, the risk of renal disease can be predicted or diagnosed as high. In addition, when the Bifidobacterium adolescentis species strain is higher than the renal disease group, it is predicted or diagnosed as a normal group, so that the renal disease risk may be predicted or diagnosed as low.

Among the biomarker strains, Bacteroides uniformis spp. strain, preferably SEQ ID NO: 7 to SEQ ID NO: 11 Bacteroides identified by any one of the rRNA nucleotide sequences of nucleotide sequence ( Bacteroides ) uniformis ) strain was significantly lower in the renal disease group than in the normal group. Therefore, when the Bacteroides uniformis species strain is lower than that of the normal group in the genomic DNA of the microorganism obtained from the sample derived from the test subject's intestinal tract, the risk of renal disease can be predicted or diagnosed as high. In addition, when the Bacteroides uniformis species strain is higher than the renal disease group, it is predicted or diagnosed as a normal group, so that the renal disease risk can be predicted or diagnosed as low.

Among the biomarker strains, Roseburia sp. strain, preferably Roseburia sp. strain identified by the 16S rRNA base sequence of SEQ ID NO: 12, is significantly higher in the renal disease group than in the normal group. . Therefore, if the genomic DNA of the microorganism obtained from the sample derived from the test subject's intestinal tract is higher than that of the normal group, it can be predicted or diagnosed with a high risk of renal disease. In addition, when the genus strain of Roseburia is lower than the renal disease group, the risk of renal disease can be predicted or diagnosed as low.

Among the biomarker strains, Oscillospira sp. strain, preferably identified by the 16S rRNA nucleotide sequence of SEQ ID NO: 13, Oscillospira sp. strain is significantly higher in the renal disease group than in the normal group. . Therefore, when the genomic DNA of the microorganism obtained from the sample derived from the test subject's intestinal tract is Oscillospira , when the spp. strain is higher than that of the normal group, the risk of renal disease can be predicted or diagnosed. In addition, Oscillospira ( Oscillospira ) When the genus strain is low compared to the renal disease group, the risk of renal disease can be predicted or diagnosed as low.

Among the biomarker strains, Lachnospiraceae and strains, preferably Lachnospiraceae identified by the 16S rRNA nucleotide sequence of SEQ ID NO: 14, are significant in the renal disease group compared to the normal group. appear low. Therefore, when the genomic DNA of the microorganism obtained from the sample derived from the test subject's intestinal tract is Lachnospiraceae and the strain is lower than that of the normal group, the risk of renal disease can be predicted or diagnosed as high. In addition, if the Lachnospiraceae and strain is higher than the renal disease group, it is predicted or diagnosed as a normal group, so that the renal disease risk can be predicted or diagnosed as low.

The 16S rRNA nucleotide sequence of SEQ ID NO: 1 to SEQ ID NO: 14 is an ASV nucleotide sequence capable of identifying each of the biomarker strains, that is, an amplicon sequence variant nucleotide sequence. In particular, the 16S rRNA nucleotide sequences of SEQ ID NOs: 1 to 4 and SEQ ID NOs: 12 to 14 are identified for the first time as nucleotide sequences capable of identifying each biomarker strain. Therefore, the biomarker strain identified by any one sequence selected from the ASV nucleotide sequence of SEQ ID NO: 1 to SEQ ID NO: 4 and SEQ ID NO: 12 to SEQ ID NO: 14 is a strain belonging to each species, genus or family, but known in the prior art These are the first strains identified in the intestine of Koreans that are clearly distinguished molecularly from strains of each species, genus or family.

In the present invention, 'risk prediction' refers to determining whether a patient is likely to develop a disease, and to delay or prevent the onset of disease through special and appropriate management of a patient with a high risk of disease, or to prevent the onset of, or the most appropriate treatment It can be used clinically to make treatment decisions by choosing a modality. In addition, 'diagnosis' means confirming the presence or characteristics of a pathological condition, and for the purpose of the present invention, diagnosis may mean confirming whether or not the onset of .

As an agent capable of detecting the strain provided as a biomarker in the present invention, a protein, nucleic acid, lipid, glycolipid, glycoprotein or sugar (monosaccharide, disaccharide, oligosaccharide, etc.) specifically present in the corresponding strain in the sample, etc. Primers, probes, antisense oligonucleotides, aptamers, antibodies, and the like capable of specifically detecting the same organic molecule may be used.

For example, when the agent for detecting the strain is a primer, it is preferable that the primer specifically detects the genomic sequence (eg, 16S rRNA) of the microorganism and does not specifically bind to the genomic sequence of another strain.

In the present invention, the term 'primer' refers to 7 to 50 nucleic acid sequences capable of forming a base pair complementary to the template strand and functioning as a starting point for copying the template strand. Primers are usually synthesized but can also be used on naturally occurring nucleic acids. The sequence of the primer does not necessarily have to be exactly the same as the sequence of the template, but is sufficiently complementary so that it can hybridize with the template. Additional features that do not change the basic properties of the primer may be incorporated. Examples of additional features that may be incorporated include, but are not limited to, methylation, encapsulation, substitution of one or more nucleic acids with homologs, and modifications between nucleic acids.

In the present invention, '16s rRNA' is an rRNA constituting the 30S subunit of the prokaryotic ribosome, and the nucleotide sequence is largely conserved, while high nucleotide sequence diversity appears in some sections. In particular, since there is little diversity among homogeneous species while diversity appears among other species, prokaryotes can be usefully identified by comparing the sequences of 16S rRNA.

In the present invention, the primer can be used to amplify the conserved 16S rRNA sequence of the microorganism, and the presence of the microorganism can be detected by whether a desired product is generated as a result of the sequence amplification. A sequence amplification method using a primer may use various methods known in the art. For example, polymerase chain reaction (PCR), reverse transcription-polymerase chain reaction (RT-PCR), multiplex PCR, touchdown PCR, hot start PCR, nested PCR, Booster PCR, real-time PCR, differential display PCR (DD-PCR), rapid amplification of cDNA ends (RACE), inverse polymerase chain reaction , vectorette PCR, TAIL-PCR (thermal asymmetric interlaced PCR), ligase chain reaction, repair chain reaction, transcription-mediated amplification, self-maintaining sequence cloning, selective amplification of the target sequence can be used, The scope of the present invention is not limited thereto.

Also, for example, when the agent for detecting the strain is an antibody, the microorganism can be detected using an immunological method based on an antigen-antibody reaction. Analysis methods for this include Western blot, ELISA (enzyme linked immunosorbent asay), radioimmunoassay (RIA), radioimmunodiffusion, Ouchterlony immunodiffusion, and rocket immunoelectrolysis. Electrophoresis, tissue immunostaining, immunoprecipitation assay, complement fixation assay, fluorescence activated cell sorter (FACS), protein chip, etc., but are not limited thereto.

In addition, molecular and immunological methods widely used in the art can be used to detect the microorganisms of the present invention.

The composition comprising the agent capable of detecting the strain of the present invention may be implemented in the form of a diagnostic kit and provided as a kit for predicting or diagnosing the risk of renal disease.

The diagnostic kit includes a primer, a probe, an antisense oligonucleotide, an aptamer, and a detection agent such as an antibody for detecting the microorganisms, as well as one or more other component compositions, solutions, or devices suitable for the analysis method. can

For example, in the present invention, a diagnostic kit including a primer specific for a corresponding microorganism may be a diagnostic kit including essential elements for performing an amplification reaction such as PCR and the like. The diagnostic kit for PCR includes a test tube or other suitable container, reaction buffer, deoxynucleotides (dNTPs), enzymes such as Taq-polymerase reverse transcriptase, DNase, RNAse inhibitors, DEPC-water, sterile water. and the like.

In the present invention, the test subject's intestinal-derived sample may be preferably a fecal sample.

In order to detect microorganisms from a sample from the intestinal tract of a test subject, general amplification techniques known in the art, for example, polymerase chain reaction, reverse transcription-polymerase chain reaction, multiplex PCR, touchdown PCR, hot start PCR, four Steed PCR, Booster PCR, Real-Time PCR, Fractional Display PCR, Rapid Amplification of cDNA Ends, Inverse PCR, Vectoret PCR, TAIL-PCR, Ligase Chain Reaction, Repair Chain Reaction, transcription-mediated amplification, self-maintaining sequence cloning, A selective amplification reaction of a target sequence may be used, but the scope of the present invention is not limited thereto.

In addition, immunological methods based on general antigen-antibody reactions known in the art, for example, Western blot, ELISA, radioimmunoassay, radioimmunodiffusion method, Oukteroni immunodiffusion method, locate immunoelectrophoresis, Tissue immunostaining, immunoprecipitation assay, complement fixation assay, FACS, protein chip, etc. may be used, but the scope of the present invention is not limited thereto.

The present invention Veillonella dispar ( Veillonella dispar ) species strain, Lachnospira ( Lachnospira ) sp. strain and Dorea formicigenerans ) In the group consisting of any one or more strains selected from the group consisting of strains An agent capable of detecting any one or more strains selected therewith, or together with the Bacteroides spp. strain, Bifidobacterium adolescentis spp. strain and Bacteroides uniformis spp. An agent capable of detecting any one or more strains selected from the group consisting of strains, or Roseburia sp. strains, Oscillospira sp. strains and Lachnospiraceae from the group consisting of strains A composition for predicting or diagnosing renal disease risk using intestinal microorganisms through a formulation capable of detecting any one or more selected strains, a kit for predicting or diagnosing renal disease risk including the same, a method for providing information for predicting or diagnosing renal disease risk, and preventing or diagnosing renal disease It can be used as a therapeutic agent screening method.

1 is a graph showing the intestinal microbial distribution of 890 Koreans.
Figure 2 shows the relative ratio distribution of all strains belonging to the Veillonella dispar species and the Veillonella dispar species identified by the ASV of SEQ ID NO: 1 in the renal disease group and the normal group; It's a boxplot.
3 is a boxplot showing the relative ratio distribution of all strains belonging to the genus Lachnospira and the ASV of SEQ ID NO: 2 in the renal disease group and normal group, and Lachnospira genus strain.
4 is a boxplot showing the relative proportion distribution of the entire strain belonging to the Dorea formicigenerans species in the renal disease group and the normal group.
5 is a boxplot showing the relative ratio distribution of all strains belonging to the genus Bacteroides and the ASV of SEQ ID NO: 4 in the renal disease group and the normal group.
6 is a box plot showing the relative proportion distribution of the entire strain belonging to the renal disease group and normal group Bifidobacterium adolescentis ( Bifidobacterium adolescentis ).
7 is a box plot showing the relative proportion distribution of the entire strain belonging to the renal disease group and normal group Bacteroides uniformis ( Bacteroides uniformis ) species.
8 is a boxplot showing the relative ratio distribution of all strains belonging to the genus Roseburia and the ASV of SEQ ID NO: 12 in the renal disease group and the normal group.
9 is a renal disease group and a normal group Oscillospira ( Oscillospira ) All strains belonging to the genus and Oscillospira identified by the ASV of SEQ ID NO: 13 It is a boxplot showing the relative proportion distribution of the genus strain.
FIG. 10 is a boxplot showing the relative ratio distribution of all strains belonging to the Lachnospiraceae family and the Lachnospiraceae family strain identified by the ASV of SEQ ID NO: 14 in the renal disease group and the normal group.
11 is a Veillonella dispar ( Veillonella dispar ) species strain of Table 1, Lachnospira spp. strain and Dorea formicigenerans ) By constructing a multivariate linear model using the relative amounts of the strain, This is a boxplot predicting the difference between the normal group and the renal disease group.
12 is a boxplot predicting the difference between a normal group and a renal disease group by constructing a multivariate linear model using the relative amounts of all biomarker microorganisms in Table 1.

Hereinafter, the present invention will be described in detail by way of experimental examples and examples. However, the following examples are merely illustrative of the present invention, and the present invention is not limited by the following examples.

Experimental Example 1: Research subject and sample collection

Fecal samples were collected from 890 Koreans participating in the health checkup. To minimize changes in fecal microorganisms, fecal samples were collected using the OMNIgene-GUT kit (DNA Genotek, Ontario, Canada) and stored at room temperature until DNA extraction.

In addition, during the health check-up, questionnaires about lifestyle, urinalysis, and blood tests were performed, and the glomerular filtration rate (eGFR) was calculated using the CKD-EPI calculation formula, taking into account the blood creatine level and the patient's age, sex, and race.

Experimental Example 2: DNA Extraction

DNA was extracted from the fecal sample of Experimental Example 1 using the bead-beating extraction method, and DNA was extracted using the QIAamp DNA Stool Mini Kit (Qiagen, Hilden, Germany).

Experimental Example 3: 16S rRNA gene sequencing

A library targeting the V3-V4 hypervariable region of the 16S rRNA gene was prepared using the DNA extracted in Experimental Example 2, and the sequence of the portion was sequenced using Illumina MiSeq 2x300 (Illumina, CA, USA).

Experimental Example 4: Sequence analysis and annotation

The sequencing result in Experimental Example 3 was converted into an amplicon sequence variant (ASV) table using the QIIME2 DADA2 module. Each ASV corresponds to a partial sequence of 16S rRNA, and each can be used as an index for detecting a specific microorganism. To identify the microbial phylogeny, the microbial phylogeny was analyzed using the Naive Bayesian classifier of QIIME2 and the GreenGene 13.8 database, and the microbial species unit annotation was performed.

Experimental Example 5: Correlation analysis between intestinal microbes and body mass index

Based on the glomerular filtration rate (eGFR) of 890 health check-up subjects, more than 90 were classified as normal and less than 90 as renal disease.

The average ratio of the total intestinal microorganisms in the renal disease group and normal group was obtained for each ASV nucleotide sequence representing a specific microbial species, and microorganisms whose average ratio sequentially increased or decreased in the renal disease group and the normal group were selected. When the ratio of the selected microorganisms showed a statistically significant difference between the renal disease group and the normal group, it was defined as a biomarker microorganism for prediction or diagnosis of renal disease risk. One-way ANOVA was used for statistical analysis, and a significant difference was shown between the renal disease group and the normal group by comparing box plots as in FIGS. 2 to 10 .

Experiment result

The distribution of intestinal microbes in 890 Koreans is shown in FIG. 1 . Intestinal microorganisms Bacteroides, Prevotella, Picalibacterium, Unclassified Lachnospiraceae, Unclassified Luminococci, Oscillospira, Luminococcus, Parabacteroides, Suterella, Coprococcus and Classified as other, the relative ratio of Bacteroides was shown by arranging from left to right from high to low subjects.

As a biomarker strain for predicting or diagnosing the risk of renal disease showing a statistically significant ratio difference between the renal disease group and the normal group, Veillonella dispar in Table 1 below, Lachnospira genus Strain, Dorea formicigenerans spp. strain, Bacteroides spp. strain, Bifidobacterium adolescentis spp. strain, Bacteroides uniformis spp. strain , Roseburia sp. strain, Oscillospira sp. strain and Lachnospiraceae and strains were selected.

division system between the renal disease group and the normal group
P-value Veillonella dispar ASV Bell 0.001944 Lachnospira ASV inside 0.006391 Dorea formicigenerans ASV Bell 0.010026 Bacteroides ASV inside 0.000850 Bifidobacterium adolescentis
( Bifidobacterium adolescentis ) Bell 0.014467 Bacteroides uniformis
( Bacteroides uniformis ) Bell 0.024214 Roseburia ASV inside 0.019845 Oscillospira ASV inside 0.020705 Lachnospiraceae ASV class 0.038335

In the renal disease group and normal group of Figure 2, Veillonella dispar ( Veillonella dispar ) Looking at the relative ratio distribution of the strain identified by the ASV of the entire strain and SEQ ID NO: 1 belonging to the Veillonella dispar ( Veillonella dispar ) species , Veillonella dispar ( Veillonella dispar ) The relative ratio of the total strains belonging to the species has no significant difference for each group, but the Veillonella dispar identified by the ASV of SEQ ID NO: 1 In the case of the strain, the normal group Compared to that, the ratio was significantly higher in the renal disease group. Therefore, if the Veillonella dispar species strain identified by the ASV of SEQ ID NO: 1 in the genomic DNA of the microorganism obtained from the sample derived from the test subject's intestinal tract is higher than that of the normal group, it can be predicted or diagnosed as having a high risk of renal disease. there is. In addition, when the Veillonella dispar species strain identified by the ASV of SEQ ID NO: 1 is low compared to the renal disease group, it corresponds to the normal group and can be predicted or diagnosed as having a low renal disease risk.

In the renal disease group and normal group of Figure 3, Lachnospira ( Lachnospira ) Looking at the relative ratio distribution of the strain identified by the ASV of SEQ ID NO: 2 and the entire strain belonging to the genus Lachnospira ( Lachnospira ), Lachnospira ( Lachnospira ) The relative ratio of the total strains belonging to the genus does not have a significant difference for each group, but Lachnospira identified by the ASV of SEQ ID NO: 2 In the case of the genus strain, it represents a significantly lower ratio in the renal disease group than in the normal group. Therefore, if the Lachnospira spp . strain identified by the ASV of SEQ ID NO: 2 in the genomic DNA of the microorganism obtained from the sample derived from the test subject's intestinal tract is lower than that of the normal group, it can be predicted or diagnosed as having a high risk of renal disease. In addition, if the Lachnospira spp . strain identified by the ASV of SEQ ID NO: 2 is higher than the renal disease group, it can be predicted or diagnosed as having a low renal disease risk corresponding to the normal group.

Looking at the relative ratio distribution of Dorea formicigenerans species strain identified by ASV of SEQ ID NO: 3 in the renal disease group and normal group of FIG. 4, Dorea formici identified by the ASV of SEQ ID NO: 3 Generans ( Dorea formicigenerans ) In the case of the strain, the renal disease group showed a significantly higher ratio than the normal group. Therefore, if the Dorea formicigenerans species strain identified by the ASV of SEQ ID NO: 3 in the genomic DNA of the microorganism obtained from the sample derived from the test subject's intestinal tract is higher than that of the normal group, the risk of renal disease is high. can In addition, when the Dorea formicigenerans species strain identified by the ASV of SEQ ID NO: 3 is lower than that of the renal disease group, it corresponds to the normal group and can be predicted or diagnosed as having a low renal disease risk. The ASV sequence of SEQ ID NO: 3 was uniquely observed in more than 15% of Koreans among the sequences of the species.

In the renal disease group and normal group of Figure 5, Bacteroides ( Bacteroides ) Looking at the relative proportion distribution of the genus Bacteroides ( Bacteroides ) identified by the ASV of SEQ ID NO: 5 and the entire strain belonging to the genus Bacteroides ( Bacteroides ) The relative ratio of the total strains belonging to the genus does not have a significant difference for each group, but the Bacteroides genus strain identified by the ASV of SEQ ID NO: 5 shows a significantly lower ratio in the renal disease group than in the normal group. Therefore, when the Bacteroides genus strain identified by the ASV of SEQ ID NO: 5 in the genomic DNA of the microorganism obtained from the test subject's intestinal-derived sample is lower than that of the normal group, the risk of renal disease can be predicted or diagnosed as high. In addition, when the Bacteroides spp. strain identified by the ASV of SEQ ID NO: 5 is higher than the renal disease group, it corresponds to the normal group and can be predicted or diagnosed as having a low renal disease risk.

6 is a renal disease group and a normal group identified by any one ASV sequence selected from the nucleotide sequence of SEQ ID NO: 5 and SEQ ID NO: 6 Bifidobacterium adolescentis ( Bifidobacterium adolescentis ) Relative ratio of all strains belonging to the species Looking at the distribution, Bifidobacterium adolescentis identified by any one ASV sequence selected from the nucleotide sequence of SEQ ID NO: 5 and SEQ ID NO: 6 For the entire strain belonging to the species, renal disease group compared to the normal group shows a significantly lower ratio in Therefore, Bifidobacterium adolescentis identified by any one ASV sequence selected from the nucleotide sequence of SEQ ID NO: 5 and SEQ ID NO: 6 in the genomic DNA of the microorganism obtained from the sample derived from the intestinal tract of the test subject All belonging to the species If the ratio of the strain is lower than that of the normal group, it can be predicted or diagnosed as having a high risk of renal disease. In addition, when the ratio of the entire strain of Bifidobacterium adolescentis identified by the ASV of SEQ ID NO: 5 and SEQ ID NO: 6 is higher than that of the renal disease group, it corresponds to the normal group and is predicted or diagnosed with a low risk of renal disease can do.

7 is a renal disease group and normal group, identified by any one ASV sequence selected from the nucleotide sequence of SEQ ID NO: 7 to SEQ ID NO: 11 Bacteroides uniformis ( Bacteroides uniformis ) Relative ratio distribution of the entire strain belonging to the species Looking at the, Bacteroides uniformis identified by any one ASV sequence selected from the nucleotide sequence of SEQ ID NO: 7 to SEQ ID NO: 11 In the case of the entire strain belonging to the species, it is significant in the renal disease group compared to the normal group shows a relatively low ratio. Therefore, in the genomic DNA of the microorganism obtained from the sample derived from the test subject's intestinal tract, Bacteroides uniformis identified by any one ASV sequence selected from the nucleotide sequences of SEQ ID NO: 7 to SEQ ID NO: 11 All strains belonging to the species If the ratio of the renal disease is lower than that of the normal group, it can be predicted or diagnosed as having a high risk of renal disease. In addition, when the ratio of the total strain of Bacteroides uniformis species identified by ASV of SEQ ID NO: 7 to SEQ ID NO: 11 is higher than that of the renal disease group, it corresponds to the normal group and is predicted or diagnosed with a low risk of renal disease. can

In the renal disease group and normal group of Figure 8, Roseburia ( Roseburia ) The entire strain belonging to the genus and Roseburia identified by the ASV of SEQ ID NO: 12 Looking at the relative ratio distribution of the genus strain, Roseburia ( Roseburia ) The relative ratio of all strains belonging to the genus does not have a significant difference for each group, but Roseburia genus strain identified by ASV of SEQ ID NO: 12 shows a significantly higher ratio in the renal disease group than in the normal group. Therefore, if the genomic DNA of the microorganism obtained from the sample derived from the test subject's intestinal tract is identified by the ASV of SEQ ID NO: 12, if the genus strain of Roseburia is higher than that of the normal group, the risk of renal disease can be predicted or diagnosed. In addition, if the Roseburia sp. strain identified by the ASV of SEQ ID NO: 12 is lower than the renal disease group, it corresponds to the normal group and can be predicted or diagnosed as having a low renal disease risk.

In the renal disease group and normal group of Figure 9, Oscillospira ( Oscillospira ) The entire strain belonging to the genus and Oscillospira identified by the ASV of SEQ ID NO: 13 Looking at the relative ratio distribution of the genus strain, Oscillospira ( Oscillospira ) The relative ratio of all strains belonging to the genus does not have a significant difference for each group, but Oscillospira identified by the ASV of SEQ ID NO: 13. In the case of the genus strain, it represents a significantly higher ratio in the renal disease group than in the normal group. Therefore, if the genomic DNA of the microorganism obtained from the sample derived from the test subject's intestinal tract is identified by the ASV of SEQ ID NO: 132, Oscillospira spp. If the genus strain is higher than that of the normal group, the risk of renal disease can be predicted or diagnosed. In addition, Oscillospira identified by the ASV of SEQ ID NO: 13 When the spp. strain is lower than the renal disease group, it corresponds to the normal group and can be predicted or diagnosed as having a low renal disease risk.

In the renal disease group and normal group of Figure 10, Lachnospiraceae ( Lachnospiraceae ) The entire strain belonging to the family and the Lachnospiraceae identified by the ASV of SEQ ID NO: 14 Looking at the relative ratio distribution of the strain, Lachnospiraceae Bird ( Lachnospiraceae ) The relative ratio of the total strains belonging to the family is not significantly different for each group, but Lachnospiraceae identified by the ASV of SEQ ID NO: 14 In the case of the Lachnospiraceae family strain, it was significantly higher in the renal disease group than in the normal group. shows a low percentage. Therefore, if the Lachnospiraceae and strain identified by the ASV of SEQ ID NO: 14 in the genomic DNA of the microorganism obtained from the sample derived from the test subject's intestinal tract is lower than the normal group, the risk of renal disease can be predicted or diagnosed. In addition, if the Lachnospiraceae strain identified by the ASV of SEQ ID NO: 14 is high compared to the renal disease group, it corresponds to the normal group and can be predicted or diagnosed as having a low renal disease risk.

Among the biomarker microorganisms of Table 1, Veillonella dispar ( Veillonella dispar ) spp. strain, Lachnospira spp. strain and Dorea formicigenerans ) Multivariate linear predicting a renal disease group by combining the strain and the Dorea formicigenerans spp. The results of predicting the difference between the normal group and the renal disease group when the model was constructed are shown in FIG. 11 . 11 is a configuration of a Bayesian Ridge model for predicting a renal disease group from the relative amounts of the three types of biomarker microorganisms. The Bayesian Ridge model is a type of linear model, and variables that have no meaning among the variables constituting the linear model are automatically excluded. way to do it In the above method, the BayesianRidge function of the Python package scikit-learn was used. According to FIG. 11 , it can be confirmed that the risk of diabetes can be predicted or diagnosed more clearly when the three types of biomarker microorganisms are combined (P value = 1.3 X 10 ^-6 ).

12 is a result of predicting renal disease risk using the Bayesian Ridge model in the same manner as in FIG. 11 using all of the biomarker microorganisms in Table 1, and the difference between the normal group and the renal disease group by combining the entire biomarker microorganisms in Table 1 It can be confirmed that it can be predicted more clearly (P value = 7.1 X 10 ^-11 ).

<110> KOREA FOOD RESEARCH INSTITUTE <120> Predicting or Diagnosing Composition for Risk of Renal Diseases Using Human Intestinal Microbiome, Diagnosing Kit, Method For Providing Information, And Screening Method For Drugs For Preventing Or Treating Renal Diseases Using The Same <130> HPC9316 <160> 14 <170> KoPatentIn 3.0 <210> 1 <211> 427 <212> DNA <213> Artificial Sequence <220> <223> Veillonella dispar ASV <400> 1 tggggaatct tccgcaatgg acgaaagtct gacggagcaa cgccgcgtga gtgatgacgg 60 ccttcgggtt gtaaagctct gttaatcggg acgaaaggcc ttcttgcgaa tagttagaag 120 gattgacggt accggaatag aaagccacgg ctaactacgt gccagcagcc gcggtaatac 180 gtaggtggca agcgttgtcc ggaattattg ggcgtaaagc gcgcgcaggc ggattggtca 240 gtctgtctta aaagttcggg gcttaacccc gtgatgggat ggaaactgcc aatctagagt 300 atcggagagg aaagtggaat tcctagtgta gcggtgaaat gcgtagatat taggaagaac 360 accagtggcg aaggcgactt tctggacgaa aactgacgct gaggcgcgaa agccagggga 420 gcgaacg 427 <210> 2 <211> 402 <212> DNA <213> Artificial Sequence <220> <223> Lachnospira ASV <400> 2 tggggaatat tgcacaatgg aggaaactct gatgcagcga cgccgcgtga gtgaagaagt 60 aattcgttat gtaaagctct atcagcaggg aagatagtga cggtacctga ctaagaagct 120 ccggctaaat acgtgccagc agccgcggta atacgtatgg agcaagcgtt atccggattt 180 actgggtgta aagggagtgt aggtggcatc acaagtcaga agtgaaagcc cggggctcaa 240 ccccgggact gcttttgaaa ctgtggagct ggagtgcagg agaggcaagt ggaattccta 300 gtgtagcggt gaaatgcgta gatattagga ggaacaccag tggcgaaggc ggcttgctgg 360 actgtaactg acactgaggc tcgaaagcgt ggggagcaaa ca 402 <210> 3 <211> 402 <212> DNA <213> Artificial Sequence <220> <223> Dorea formicigenerans ASV <400> 3 tggggaatat tgcacaatgg gcgaaagcct gatgcagcga cgccgcgtga aggatgaagt 60 atttcggtat gtaaacttct atcagcaggg aagaaaatga cggtacctga ctaagaagcc 120 ccggctaact acgtgccagc agccgcggta atacgtaggg ggcaagcgtt atccggattt 180 actgggtgta aagggagcgt agacggctgt gcaagtctga agtgaaaggc atgggctcaa 240 cctgtggact gctttggaaa ctgtgcagct agagtgtcgg agaggtaagt ggaattccta 300 gtgtagcggt gaaatgcgta gatattagga ggaacaccag tggcgaaggc ggcttactgg 360 acgatgactg acgttgaggc tcgaaagcgt ggggagcaaa ca 402 <210> 4 <211> 422 <212> DNA <213> Artificial Sequence <220> <223> Bacteroides ASV <400> 4 tgaggaatat tggtcaatgg gcgcaggcct gaaccagcca agtagcgtga aggatgactg 60 ccctatgggt tgtaaacttc ttttataaag gaataaagtc gggtatgtat acccgtttgc 120 atgtacttta tgaataagga tcggctaact ccgtgccagc agccgcggta atacggagga 180 tccgagcgtt atccggattt attgggttta aagggagcgt agatggatgt ttaagtcagt 240 tgtgaaagtt tgcggctcaa ccgtaaaatt gcagttgata ctggatatct tgagtgcagt 300 tgaggcaggc ggaattcgtg gtgtagcggt gaaatgctta gatatcacga agaactccga 360 ttgcgaaggc agcctgctaa gctgcaactg acattgaggc tcgaaagtgt gggtatcaaa 420 ca 422 <210> 5 <211> 411 <212> DNA <213> Artificial Sequence <220> <223> Bifidobacterium adolescentis <400> 5 tggggaatat tgcacaatgg gcgcaagcct gatgcagcga cgccgcgtgc gggatgacgg 60 ccttcgggtt gtaaaccgct tttgactggg agcaagccct tcggggtgag tgtacctttc 120 gaataagcac cggctaacta cgtgccagca gccgcggtaa tacgtagggt gcaagcgtta 180 tccggaatta ttgggcgtaa agggctcgta ggcggttcgt cgcgtccggt gtgaaagtcc 240 atcgcttaac ggtggatccg cgccgggtac gggcgggctt gagtgcggta ggggagactg 300 gaattcccgg tgtaacggtg gaatgtgtag atatcgggaa gaacaccaat ggcgaaggca 360 ggtctctggg ccgttactga cgctgaggag cgaaagcgtg gggagcgaac a 411 <210> 6 <211> 411 <212> DNA <213> Artificial Sequence <220> <223> Bifidobacterium adolescentis <400> 6 tggggaatat tgcacaatgg gcgcaagcct gatgcagcga cgccgcgtgc gggatgacgg 60 ccttcgggtt gtaaaccgct tttgactggg agcaagccct tcggggtgag tgtacctttc 120 gaataagcac cggctaacta cgtgccagca gccgcggtaa tacgtagggt gcaagcgtta 180 tccggaatta ttgggcgtaa agggctcgta ggcggttcgt cgcgtccggt gtgaaagtcc 240 atcgcttaac ggtggatccg cgccgggtac gggcgggctt gagtgcggta ggggagactg 300 gaattcccgg tgtaacggtg gaatgtgtag atatcgggaa gaacaccaat ggcgaaggca 360 ggtctctggg ccgtcactga cgctgaggag cgaaagcgtg gggagcgaac a 411 <210> 7 <211> 422 <212> DNA <213> Artificial Sequence <220> <223> Bacteroides uniformis <400> 7 tgaggaatat tggtcaatgg acgagagtct gaaccagcca agtagcgtga aggatgactg 60 ccctatgggt tgtaaacttc ttttatacgg gaataaagtg aggcacgcgt gcctttttgt 120 atgtaccgta tgaataagga tcggctaact ccgtgccagc agccgcggta atacggagga 180 tccgagcgtt atccggattt attgggttta aagggagcgt aggcggacgc ttaagtcagt 240 tgtgaaagtt tgcggctcaa ccgtaaaatt gcagttgata ctgggtgtct tgagtacagt 300 agaggcaggc ggaattcgtg gtgtagcggt gaaatgctta gatatcacga agaactccga 360 ttgcgaaggc agcctgctgg actgtaactg acgctgatgc tcgaaagtgt gggtatcaaa 420 ca 422 <210> 8 <211> 422 <212> DNA <213> Artificial Sequence <220> <223> Bacteroides uniformis <400> 8 tgaggaatat tggtcaatgg acgagagtct gaaccagcca agtagcgtga aggatgactg 60 ccctatgggt tgtaaacttc ttttatacgg gaataaagtg aggcacgcgt gcctttttgt 120 atgtaccgta tgaataagga tcggctaact ccgtgccagc agccgcggta atacggagga 180 tccgagcgtt atccggattt attgggttta aagggagcgt aggcggacgc ttaagtcagt 240 tgtgaaagtt tgcggctcaa ccgtaaaatt gcagttgata ctgggtgtct tgagtacagt 300 agaggcaggc ggaattcgtg gtgtagcggt gaaatgctta gatatcacga agaactccga 360 ttgcgaaggc agcttgctgg actgtaactg acgctgatgc tcgaaagtgt gggtatcaaa 420 ca 422 <210> 9 <211> 422 <212> DNA <213> Artificial Sequence <220> <223> Bacteroides uniformis <400> 9 tgaggaatat tggtcaatgg acgagagtct gaaccagcca agtagcgtga aggatgactg 60 ccctatgggt tgtaaacttc ttttatacgg gaataaagtg aggcacgtgt gcctttttgt 120 atgtaccgta tgaataagga tcggctaact ccgtgccagc agccgcggta atacggagga 180 tccgagcgtt atccggattt attgggttta aagggagcgt aggcggacgc ttaagtcagt 240 tgtgaaagtt tgcggctcaa ccgtaaaatt gcagttgata ctgggtgtct tgagtacagt 300 agaggcaggc ggaattcgtg gtgtagcggt gaaatgctta gatatcacga agaactccga 360 ttgcgaaggc agcttgctgg actgtaactg acgctgatgc tcgaaagtgt gggtatcaaa 420 ca 422 <210> 10 <211> 422 <212> DNA <213> Artificial Sequence <220> <223> Bacteroides uniformis <400> 10 tgaggaatat tggtcaatgg acgagagtct gaaccagcca agtagcgtga aggatgactg 60 ccctatgggt tgtaaacttc ttttatacgg gaataaagtg aggcacgtgt gcctttttgt 120 atgtaccgta tgaataagga tcggctaact ccgtgccagc agccgcggta atacggagga 180 tccgagcgtt atccggattt attgggttta aagggagcgt aggcggacgc ttaagtcagt 240 tgtgaaagtt tgcggctcaa ccgtaaaatt gcagttgata ctgggtgtct tgagtacagt 300 agaggcaggc ggaattcgtg gtgtagcggt gaaatgctta gatatcacga agaactccga 360 ttgcgaaggc agcctgctgg actgtaactg acgctgatgc tcgaaagtgt gggtatcaaa 420 ca 422 <210> 11 <211> 422 <212> DNA <213> Artificial Sequence <220> <223> Bacteroides uniformis <400> 11 tgaggaatat tggtcaatgg acgagagtct gaaccagcca agtagcgtga aggatgactg 60 ccctatgggt tgtaaacttc ttttatacgg gaataaagtt aggcacgtgt gcctttttgt 120 atgtaccgta tgaataagga tcggctaact ccgtgccagc agccgcggta atacggagga 180 tccgagcgtt atccggattt attgggttta aagggagcgt aggcggatgc ttaagtcagt 240 tgtgaaagtt tgcggctcaa ccgtaaaatt gcagttgata ctgggtgtct tgagtacagt 300 agaggcaggc ggaattcgtg gtgtagcggt gaaatgctta gatatcacga agaactccga 360 ttgcgaaggc agcttgctgg actgtaactg acgctgatgc tcgaaagtgt gggtatcaaa 420 ca 422 <210> 12 <211> 404 <212> DNA <213> Artificial Sequence <220> <223> Roseburia ASV <400> 12 tggggaatat tgcacaatgg gggaaaccct gatgcagcga cgccgcgtga gcgaagaagt 60 atttcggtat gtaaagctct atcagcaggg aagaagaaat gacggtacct gactaagaag 120 caccggctaa atacgtgcca gcagccgcgg taatacgtat ggtgcaagcg ttatccggat 180 ttactgggtg taaagggagc gcaggcggaa ggctaagtct gatgtgaaag cccggggctc 240 aaccccggta ctgcattgga aactggtcat ctagagtgtc ggaggggtaa gtggaattcc 300 tagtgtagcg gtgaaatgcg tagatattag gaggaacacc agtggcgaag gcggcttact 360 ggacgataac tgacgctgag gctcgaaagc gtggggagca aaca 404 <210> 13 <211> 404 <212> DNA <213> Artificial Sequence <220> <223> Oscillospira ASV <400> 13 tggggaatat tgggcaatgg gcgcaagcct gacccagcaa cgccgcgtga aggaagaagg 60 ctttcgggtt gtaaacttct tttaagaggg aagagcagaa gacggtacct ctagaataag 120 ccacggctaa ctacgtgcca gcagccgcgg taatacgtag gtggcaagcg ttgtccggat 180 ttactgggtg taaagggcgt gcagccgggt ctgcaagtca gatgtgaaat ccatgggctc 240 aacccatgaa ctgcatttga aactgtagat cttgagtgtc ggaggggcaa tcggaattcc 300 tagtgtagcg gtgaaatgcg tagatattag gaggaacacc agtggcgaag gcggattgct 360 ggacgataac tgacggtgag gcgcgaaagt gtggggagca aaca 404 <210> 14 <211> 402 <212> DNA <213> Artificial Sequence <220> <223> Lachnospiraceae ASV <400> 14 tggggaatat tgcacaatgg gcgaaagcct gatgcagcaa cgccgcgtga gtgaagaagt 60 atctcggtat gtaaagctct atcagcaggg aagaaaatga cggtacctga ctaagaagcc 120 ccggctaact acgtgccagc agccgcggta atacgtaggg ggcaagcgtt atccggattt 180 actgggtgta aagggagcgc agacggcact gcaagtctga agtgaaagcc cggggctcaa 240 ccccgggact gctttggaaa ctgtagagct agagtgctgg agaggcaagc ggaattccta 300 gtgtagcggt gaaatgcgta gatattagga agaacaccag tggcgaaggc ggcttgctgg 360 acagtaactg acgttcaggc tcgaaagcgt ggggagcaaa ca 402

Claims (17)

Lachnospira ( Lachnospira ) sp. strain and Dorea formicigenerans ( Dorea formicigenerans ) For predicting or diagnosing renal disease risk using intestinal microorganisms, including an agent capable of detecting any one or more strains selected from the group consisting of strains composition. According to claim 1, wherein the Lachnospira ( Lachnospira ) genus strain is to include the 16S rRNA base sequence of SEQ ID NO: 2, The Dorea formicigenerans species strain is 16S rRNA base of SEQ ID NO: 3 A composition for predicting or diagnosing the risk of renal disease using an intestinal microorganism, characterized in that it comprises a sequence. According to claim 1, wherein the Lachnospira ( Lachnospira ) sp. strain and Dorea formicigenerans ) In the agent capable of detecting any one or more strains selected from the group consisting of strains, Bacteroides ( Bacteroides ) sp. strain, Bifidobacterium adolescentis spp. strain, and Bacteroides uniformis ) An agent capable of detecting any one or more strains selected from the group consisting of strains further Including, a composition for predicting or diagnosing the risk of renal disease using intestinal microorganisms. According to claim 3, wherein the Bacteroides ( Bacteroides ) sp. strain is to include the 16S rRNA base sequence of SEQ ID NO: 4, the Bifidobacterium adolescentis ( Bifidobacterium adolescentis ) The strain is SEQ ID NO: 5 and SEQ ID NO: It will include any one sequence selected from the 16S rRNA nucleotide sequence of 6, and the Bacteroides uniformis species strain is any one selected from the 16S rRNA nucleotide sequence of SEQ ID NO: 7 to SEQ ID NO: 11 A composition for predicting or diagnosing the risk of renal disease using an intestinal microorganism, characterized in that it comprises a sequence. According to claim 3, Roseburia ( Roseburia ) sp. strain, Oscillospira sp. strain and Lachnospiraceae ( Lachnospiraceae ) An agent capable of detecting any one or more strains selected from the group consisting of strains Further comprising, a composition for predicting or diagnosing the risk of renal disease using intestinal microorganisms. According to claim 5, wherein the Roseburia ( Roseburia ) sp. strain is to include the 16S rRNA nucleotide sequence of SEQ ID NO: 12, the Oscillospira ( Oscillospira ) sp. strain comprising the 16S rRNA nucleotide sequence of SEQ ID NO: 13 The Lachnospiraceae and strain is a composition for predicting or diagnosing renal disease risk using an intestinal microorganism, characterized in that it contains the 16S rRNA base sequence of SEQ ID NO: 14. The renal disease using intestinal microorganisms according to any one of claims 1 to 6, wherein the agent capable of detecting the strain is a primer, probe, antisense oligonucleotide, aptamer or antibody specific to the strain. A composition for risk prediction or diagnosis. The composition for predicting or diagnosing the risk of renal disease using an intestinal microorganism according to claim 7, wherein the primer is a primer capable of amplifying the 16S rRNA of the strain. A kit for predicting or diagnosing renal disease risk, comprising the composition of any one of claims 1 to 6. Detecting any one or more strains selected from the group consisting of Lachnospira sp . strains and Dorea formicigenerans sp. strains from the genomic DNA of microorganisms obtained from the intestinal-derived sample of the test subject; and
The Lachnospira ( Lachnospira ) sp. strain and Dorea formicigenerans ( Dorea formicigenerans ) comparing the increase or decrease of any one or more strains selected from the group consisting of strains with the normal group; renal disease using intestinal microorganisms comprising; As a method for providing information necessary for risk prediction or diagnosis,
The renal disease or renal disease risk group, characterized in that the following selection criteria are satisfied:
Compared to the normal group, the genomic DNA of the test subject's microorganisms Lachnospira ( Lachnospira ) that the spp. strain is reduced, or
In the genomic DNA of the test subject's microorganism compared to the normal group, the Dorea formicigenerans species strain is increased. 11. The method of claim 10, wherein from the genomic DNA of the microorganism obtained from the sample derived from the intestinal tract of the subject Bacteroides spp. strain, Bifidobacterium adolescentis spp. strain and Bacteroides uniformis ( Bacteroides uniformis ) further detecting any one or more strains selected from the group consisting of species strains; and
The Bacteroides ( Bacteroides ) sp. strain, Bifidobacterium adolescentis ( Bifidobacterium adolescentis ) Sp. strain and Bacteroides uniformis ( Bacteroides uniformis ) Normal increase or decrease of any one or more strains selected from the group consisting of strains Comparing with the group; further comprising,
The renal disease or renal disease risk group, characterized in that the following selection criteria are satisfied:
Compared to the normal group, in the genomic DNA of the test subject's microorganism, the Bacteroides spp. strain is reduced,
Bifidobacterium adolescentis in the genomic DNA of the microorganisms of the test subject compared to the normal group that the strain is reduced, or
Compared to the normal group, the Bacteroides uniformis species strain is reduced in the genomic DNA of the test subject's microorganism. 12. The method of claim 11, wherein from the genomic DNA of the microorganism obtained from the sample derived from the intestinal tract of the test subject, Roseburia sp. strain, Oscillospira sp. strain and Lachnospiraceae group consisting of strains. Further detecting any one or more strains selected from; and
The Roseburia ( Roseburia ) sp. strain, Oscillospira sp. strain and Lachnospiraceae ( Lachnospiraceae ) Comparing the increase or decrease of any one or more strains selected from the group consisting of strains with the normal group; including as
The renal disease or renal disease risk group, characterized in that the following selection criteria are satisfied:
Compared to the normal group, in the genomic DNA of the test subject's microorganism, Roseburia spp . strain is increased,
Oscillospira in the genomic DNA of the test subject's microorganism compared to the normal group, the genus strain is increased, or
Compared to the normal group, Lachnospiraceae and strains in the genomic DNA of the test subject's microorganisms are reduced. From the genomic DNA of microorganisms obtained from the sample derived from the intestinal tract of the test subject, Lachnospira sp. strain, Dorea formicigenerans sp. strain, Bacteroides sp. strain, Bifidobacterium adoles Select from Bifidobacterium adolescentis spp. strains and Bacteroides uniformis spp. strains, Roseburia sp. strains, Oscillospira sp. strains and Lachnospiraceae family strains detecting the relative amounts of the two or more strains; and
The result obtained in the detecting step, calculating the renal disease risk of the test subject by a multivariate linear model predicting a renal disease group using the relative amount of the two or more strains as a variable; Information provision method for risk prediction or diagnosis. From the genomic DNA of microorganisms obtained from the sample derived from the intestinal tract of the test subject, Lachnospira sp. strain, Dorea formicigenerans sp. strain, Bacteroides sp. strain, Bifidobacterium adoles Centis ( Bifidobacterium adolescentis ) Species strains and Bacteroides uniformis spp. strains, Roseburia genus strains, Oscillospira genus strains and Lachnospiraceae family strains detected to do; and
The Lachnospira ( Lachnospira ) sp. strain, Dorea formicigenerans spp. strain, Bacteroides sp. strain, Bifidobacterium adolescentis spp. strain and Bacteroides uni Formis ( Bacteroides uniformis ) species strain, Roseburia spp. strain, Oscillospira spp. strain and Lachnospiraceae ) Comparing the increase and decrease of the family strain with the normal group; further comprising; and,
The renal disease or renal disease risk group, characterized in that the following selection criteria are satisfied:
Compared to the normal group, in the genomic DNA of the test subject's microorganisms, the Dorea formicigenerans spp. strain, Roseburia spp . strain or Oscillospira spp. strain is increased, or
Compared to the normal group, in the genomic DNA of the microorganisms of the test subject, Lachnospira sp. strain, Bacteroides sp. strain, Bifidobacterium adolescentis spp. strain, Bacteroides uniformis ( Bacteroides uniformis ) Species strains or Lachnospiraceae and those with reduced strains. administering a candidate substance for the prevention or treatment of renal disease to a non-human animal;
Any one or more strains selected from the group consisting of Lachnospira sp . strains and Dorea formicigenerans sp. strains from the genomic DNA of microorganisms obtained from the intestinal-derived sample before and after the candidate material treatment detecting; and
Comparing the increase or decrease of any one or more strains selected from the group consisting of Lachnospira sp. strains and Dorea formicigenerans strains in the genomic DNA of microorganisms before and after treatment with the candidate material, Veillonella dispar ( Veillonella dispar ) When the strain is reduced, or Lachnospira ( Lachnospira ) When the spp. strain is increased, or Dorea formicigenerans ) When the strain is reduced A renal disease prevention or therapeutic agent screening method using intestinal microorganisms comprising; determining the candidate substance as a renal disease prevention or therapeutic agent. According to claim 15, Bacteroides ( Bacteroides ) sp. strain, Bifidobacterium adolescentis ( Bifidobacterium adolescentis ) sp. strain and Bacteroides from the genomic DNA of microorganisms obtained from the sample derived from the intestinal tract before and after the candidate material treatment further detecting any one or more strains selected from the group consisting of Bacteroides uniformis strains; and
Bacteroides spp. strain, Bifidobacterium adolescentis spp. strain and Bacteroides uniformis spp. strain in the genomic DNA of the microorganism before and after the candidate material treatment Comparing the increase or decrease of any one or more strains selected from the group, Bacteroides ( Bacteroides ) When the spp. strain is increased, or Bifidobacterium adolescentis ( Bifidobacterium adolescentis ) When the strain is increased, or Bac Teroides uniformis ( Bacteroides uniformis ) In case the strain is increased, determining the candidate material as a renal disease prevention or treatment agent; 17. The method of claim 16, wherein from the genomic DNA of the microorganism obtained from the sample derived from the intestinal tract before and after treatment with the candidate material, a spp . Further detecting any one or more strains selected from the group consisting of strains; and
In the genomic DNA of the microorganism before and after the candidate material treatment, Roseburia sp. strain, Oscillospira sp. strain and Lachnospiraceae Any one or more strains selected from the group consisting of strains Comparing the increase or decrease of, Roseburia ( Roseburia ) If the strain is reduced, or if the oscillospira ( Oscillospira ) sp. strain is reduced, or Lachnospiraceae ( Lachnospiraceae ) and the strain is increased A renal disease prevention or therapeutic agent screening method using intestinal microorganisms comprising; determining the candidate substance as a renal disease prevention or therapeutic agent.

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